[Acute renal failure in hantavirus infections]. / Akutes Nierenversagen bei Hantavirusinfektionen.

BACKGROUND: The acute renal failure remains a diagnostic challenge for the clinician. CASE REPORTS: Between 1991 and 1996, acute renal failure caused by hantavirus infection was diagnosed in 4 previously healthy male patients. Main symptoms consisted of fever, headache, arthralgia, lumbar and abdominal pain as well as a decline in diuresis. The ultrasonography showed a slight splenomegaly in 2 patients. The clinical chemistry showed elevated serum creatinine from 2.2 mg/dl to 6.7 mg/dl and thrombocytopenia from 4000 to 150,000/microliter. The examination of the urine showed slight proteinuria and microhematuria. The kidney biopsy of 1 patient showed a reversible damage of the tubuli. The pathologic findings normalized within 3 weeks in 3 patients without need for dialysis. One patient developed a severe clinical course with acute renal failure and pulmonary edema requiring dialysis. In all patients, the renal function improved.

A new Clethrionomys-derived hantavirus from Germany: evidence for distinct genetic sublineages of Puumala viruses in Western Europe.

Puumala (PUU) viruses are the predominant etiologic agents of hantavirus infections in Europe. The most important reservoir is the bank vole, Clethrionomys glareolus (Cg), belonging to the subfamily Arvicolinae of the Muridae family. Here we report on the molecular characterization of the first rodent-derived sequence (PUU/Cg-Erft) from Germany. Comparison of the S and M segment coding regions revealed 92.5 and 92.8% identity, respectively, with PUU/H-9013, a human isolate from France. However, only 83.1% identity was found with the S segment of a previously reported PUU sequence from a German HFRS case (PUU/H-Berkel) indicating the co-existence of two distinct sublineages in Germany. Phylogenetic and alignment analyses of S and M segment coding regions enabled us to assign PUU viruses/sequences to at least six distinct genetic sublineages. Membership was defined by nucleotide sequence differences of < 8%, whereas a diversity of > 14% clearly outgrouped a virus/sequence. Based on S segment sequences the sublineage represented by Clethrionomys rufocanus-derived viruses from Japan diverged at a well supported node from the clade harbouring all Clethrionomys glareolus-derived European PUU viruses. A correlation between genetic relationship and geographic origin of PUU viruses was observed which may support a co-evolution of PUU viruses with distinct subspecies of their reservoir host.

Polymerase chain reaction detection of Puumala virus RNA in formaldehyde-fixed biopsy material.

BACKGROUND: Infections with hantaviruses, mainly Clethrionomys-derived Puumala viruses, are known causes of acute renal failure [hemorrhagic fever with renal syndrome (HFRS)] in western Europe. Laboratory diagnosis is primarily based on serology. At the time of clinical symptoms, viral RNA can hardly be detected in the blood or urine, indicating that polymerase chain reaction (PCR) is of little diagnostic value for these infections. Biopsy material is usually formaldehyde-fixed and, thus, regarded as poor quality for PCR applications. The aim of this study was to establish a technique to retrieve such material for laboratory diagnostic. METHODS: Formaldehyde-fixed, paraffin-embedded kidney biopsies of 14 patients with renal failure either clinically suspected for HFRS (7 cases) or caused by unknown (2 cases) or known other causes (drugs, sarcoidosis; 5 cases) were histologically investigated. An established S segment-specific PCR assay was applied to RNA isolated from the biopsies, and amplification products were verified by direct sequence determination. RESULTS: Investigations revealed a typical histopathological appearance for hantavirus infections in all seven suspected HFRS cases and one case of unknown cause. With five of the suspected HFRS cases, hantavirus-specific RNA was detected. Sequence comparison revealed a close relationship to corresponding nucleoproteins of known Puumala viruses. CONCLUSION: The established technique provides a simple and powerful tool that expands the diagnostic possibilities, especially for otherwise unidentified or retrospective cases. It further allows insight into the molecular epidemiology of HFRS-causing agents.

[Acute hantavirus infection caused by a genetically newly identified viral strain. Severe and complicated course of hemorrhagic fever with renal syndrome (nephropathia epidemica)]. / Akute Hantavirusinfektion durch genetisch neu identifizierten Virusstamm. Schwerer und komplizierter Verlauf eines hämorrhagischen Fiebers mit renalem Syndrom (Nephropathia epidemica).

HISTORY AND CLINICAL FINDINGS: A 49-year-old patient, a hobby hunter, fell ill acutely with joint and limb pain, abdominal pain, nausea and subfebrile temperatures. At hospitalization, the patient was in bad general condition, showing ascites and lid edema, and acute renal failure was diagnosed. INVESTIGATIONS: Laboratory tests revealed marked thrombocytopenia (15,000/ml), leucocytosis, elevated levels of creatinine, blood urea nitrogen and liver enzymes, respectively. Blood gas analysis showed metabolic acidosis. Chest X-ray showed an interstitial fluid accumulation, abdominal ultrasound disclosed ascites and enlarged kidneys as in acute renal failure. Immunologic tests verified the diagnosis of an acute hantavirus infection, by use of specific molecular biology techniques a previously unknown virus strain was identified. TREATMENT AND COURSE: Hantavirus infections in western Europe usually show a benign course. However, in the present case, acute progressive pulmonary failure developed despite effective dialysis so that mechanical ventilation was necessary for several weeks. Dialysis had to be carried out for 17 days. As a complication a severe ulcero-destructive tracheobronchitis developed, caused by Aspergillus fumigatus. After several weeks, both renal and pulmonary function had returned to normal. CONCLUSION: Hantavirus infections may lead to severe and complicated courses also in western Europe. By use of new immunologic and molecular biology techniques a specific diagnosis is possible.

Genetic identification of a new Puumala virus strain causing severe hemorrhagic fever with renal syndrome in Germany.

A severe case of suspected hemorrhagic fever with renal syndrome (HFRS) was recently identified in northwestern Germany. A genetic detection assay was designed that identified hantavirus-specific RNA in the patient's clinical specimens by reverse transcriptase-polymerase chain reaction amplification of virus S and M genome segments. Phylogenetic analysis of the nucleotide sequences demonstrated that this virus belonged to the Puumala (PUU) group, with the closest relationship to a PUU isolate from Finland. Within the group, this virus formed a separate lineage. This finding represents the first genetic characterization of a hantavirus causing severe HFRS in Germany. The data suggest that PUU viruses circulating in western European countries are genetically distinct from their northeastern counterparts. Comparison of deduced amino acid sequences demonstrated a loss of a potential N-glycosylation site in the G2 protein compared with other PUU viruses.

Genomic characterization of a poxvirus isolated from a child.

A poxvirus was isolated from a six-year-old girl. The comparative analyses of the genome of this isolate (H-CP-LSax) which were carried out using the restriction endonucleases BamHI, HindIII, KpnI, MluI, NcoI, SacI, and SmaI revealed that this isolate is a member of the genus orthopoxvirus. Since the girl had never been vaccinated against smallpox, and had close contact to domestic animals, including cats, rabbits and guinea pigs, the genome of H-CP-LSax virus was genetically analysed in comparison with other known orthopoxviruses. The analysis demonstrates clearly that the HindIII cleavage pattern of H-CP-LSax DNA is different from the HindIII DNA cleavage patterns of vaccinia virus, cowpox virus, rabbit poxvirus, cat poxvirus, ectromelia virus, and okapi poxvirus. Surprisingly, it was found that the HindIII and SmaI cleavage patterns of the DNA of one out of six elephant poxviruses which were analysed under the same conditions were virtually identical to the HindIII and SmaI cleavage patterns of H-CP-LSax DNA. Although SmaI and HindIII digestion of both virus genomes gave the same fragment patterns, the viral DNAs can be distinguished from each other by the restriction endonucleases SacI, BamHI, and KpnI, which also show high similarities in the fragmentation patterns of both viruses. The results obtained in this study indicate three possibilities concerning the origin of H-CP-LSax virus. Firstly that the H-CP-LSax virus originated from an unknown animal species. Secondly, that this virus is a variant of elephant poxvirus in which the HindIII and SmaI sites are extremely conserved, and finally that H-CP-LSax can be a recombinant virus of unknown origin.

Comparative analysis of the genomes of orthopoxviruses isolated from elephant, rhinoceros, and okapi by restriction enzymes. Brief report.

Orthopoxviruses from different zoo-kept mammalian species including Elephas maximus (8 isolates), Ceratotherium simum (1 isolate), and Okapia johnstoni (2 isolates) were characterized by restriction enzyme analysis of the viral genome. The four enzymes BamHI, MluI, NcoI, and SalI were found to be optimal for strain differentiation.

Human antibody response to immunization with 17D yellow fever and inactivated TBE vaccine.

The antibody response against flaviviruses tick-borne encephalitis (TBE), Kyasanur Forest disease (KFD), Murray Valley encephalitis (MVE), West Nile fever (WNF), Japanese B encephalitis (JE), dengue 2 (DEN-2), and yellow fever (YF) was studied in humans after administration of an inactivated TBE virus vaccine. Individuals were either prevaccinated with 17D yellow fever (experimental group) or without any previous exposure to flaviviruses (control group). The appearance of serum titres of homologous and heterologous haemagglutination inhibition (HI) antibodies, heterotypic DEN-2 neutralizing antibodies, and TBE enzyme-linked immunosorbent assay (ELISA) antibodies were examined. Individuals prevaccinated with the 17D yellow fever developed an antibody pattern that contrasted with that of the control group. This pattern was characterized as follows: (1) Predominantly anti-TBE IgG antibodies appeared earlier and in higher titres than in the control group, (2) heterologous HI antibodies cross-reacting with the WN flavivirus subgroup preceded the appearance of homologous HI antibodies, (3) a broad spectrum HI response was observed against all flaviviruses tested, and (4) low titre heterotypic DEN-2 neutralizing antibodies were formed in about half of the cases. These observations are discussed in the context of cross-reactivity, cross-protection and virus infection enhancement.

[Isolation of Tahyna virus from mosquitoes in 2 different European natural foci]. / Nachweis des Tahyna-Virus bei Stechmücken in zwei verschiedenen europäischen Naturherden.

Collecting during two periods, from September 4, 1979, to September 12, 1980, and from August 17 to September 10, 1981, a total of 45,705 mosquitoes was caught for virus isolation studies in 6 different regions in Germany and the Netherlands (Lower Rhine area, Upper Rhine area near Germersheim, Upper Main area near Baunach, Amper Moos), in Austria (eastern shore of Lake Neusiedl), and in Italy (Isonzo river delta). 25 mosquito species were identified belonging to the genera Aedes, Culex, Culiseta, Mansonia, and Uranotaenia. The relative mosquito species composition was determined for the different collecting sites. By intracerebral inoculation of 2 to 4 days old suckling mice with extracts of each mosquito pool 6 virus strains were isolated which were identified as Tahyna (TAH) virus strains using the indirect immunofluorescence technique in cell culture and the baby mouse neutralization test. The origin of these strains were the following regions: one TAH virus strain was isolated from Aedes caspius mosquitoes (1 of 158 pools) collected from 17. 8 to 30. 8. 81 at the eastern shore of Lake Neusiedl. 5 additional isolates were obtained from 6,066 mosquitoes (62 pools) collected on 9. and 10. 9. 81 in the Upper Rhine area near Germersheim (Isle Grün). Two of these virus strains were isolated from Aedes vexans and three from unidentified mosquitoes. This is the first TAH virus isolation from mosquitoes in the Upper Rhine area, which had been regarded by Ackermann and coworkers (1970) and Spieckermann and Ackermann (1974) as a potential natural TAH virus focus on the basis of serological studies in humans and sentinel rabbits during 1969. The isolation of TAH virus in the Lake Neusiedl area essentially confirms the results obtained by Aspöck and Kunz (1967) for this region. The possible influence of some ecological factors on the geographical distribution of TAH virus is discussed.

Investigation on the thermostability of steroid hormone receptors in lyophilized calf uterine tissue powder.

Powdered calf uteri were lyophilized and sealed in a vacuum. The effects of lyophilization and the thermostability of the binding activity of uterine steroid hormone receptors were studied. The lyophilized powder was analyzed for its ability to bind estrogen, progestin, androgen, and glucocorticoid steroids. Lyophilization had no deleterious effects on the binding activity. Storage of the lyophilized powder at +20 C for more than 14 days did not reduce the binding activity. No reduction of the binding capacity was also observed when the storage temperature was at +30 C for 16 hours. However, a slight loss of specific binding sites was observed for 17 beta-estradiol (-12%) at the storage temperature of +40 C, but the other steroid binding proteins were more stable. When the storage temperature was at +60 C for 16 hours, a loss of about 24% was observed for the specific estrogen binding sites, whereas the binding capacity of progestin, androgen, and glucocorticoid receptors was reduced by only less than 10%. Storage at the high temperature of +80 C for 16 hours resulted in a loss of only 36% for the estrogen binding capacity and less than 30% loss for the other binding proteins. The freeze-dried uterine powders can be useful quality control specimens for steroid receptor assays.